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Alkylation of Staurosporine to Derive a Kinase Probe for Fluorescence Applications.

ABSTRACT: The natural product staurosporine is a high-affinity inhibitor of nearly all mammalian protein kinases. The labelling of staurosporine has proven effective as a means of generating protein kinase research tools. Most tools have been generated by acylation of the 4'-methylamine of the sugar moiety of staurosporine. Herein we describe the alkylation of this group as a first step to generate a fluorescently labelled staurosporine. Following alkylation, a polyethylene glycol linker was installed, allowing subsequent attachment of fluorescein. We report that this fluorescein-staurosporine conjugate binds to cAMP-dependent protein kinase in the nanomolar range. Furthermore, its binding can be antagonised with unmodified staurosporine as well as ATP, indicating it targets the ATP binding site in a similar fashion to native staurosporine. This reagent has potential application as a screening tool for protein kinases of interest.

SUBMITTER: Disney AJ

PROVIDER: S-EPMC4949516 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature

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Alkylation of Staurosporine to Derive a Kinase Probe for Fluorescence Applications.

Disney Alexander J M AJ Kellam Barrie B Dekker Lodewijk V LV

ChemMedChem 20160323 9

The natural product staurosporine is a high-affinity inhibitor of nearly all mammalian protein kinases. The labelling of staurosporine has proven effective as a means of generating protein kinase research tools. Most tools have been generated by acylation of the 4'-methylamine of the sugar moiety of staurosporine. Herein we describe the alkylation of this group as a first step to generate a fluorescently labelled staurosporine. Following alkylation, a polyethylene glycol linker was installed, al ...[more]

PMID: 27008372

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